title: Interaction of lipoprotein lipase with subendothelial extracellular matrix.
abstract: We have analyzed the binding of lipoprotein lipase (LPL) to the subendothelial extracellular matrix produced by cultured endothelial cells. Binding was linear up to a concentration of 0.5 microgram/ml (10 nM) enzyme used in this study, and equilibrium was achieved after 2 h of incubation with bovine 125I-LPL at 4 degrees C. Heparin and heparan sulfate effectively inhibited the binding of LPL to extracellular-matrix-coated plates; chondroitin sulfate had no effect, while high concentrations of dermatan sulfate or keratan sulfate inhibited binding of LPL to extracellular matrix by only 40%. Basic fibroblast growth factor (bFGF) did not affect LPL binding, while antithrombin-III (AT-III) caused up to a 50% inhibition of enzyme binding to extracellular matrix. alpha-Thrombin. 5.10(-6) M, and its esterolytically inactive derivative, DIP-alpha-thrombin, effectively inhibited binding of LPL to extracellular-matrix-coated plates. alpha-Thrombin was also able to release the extracellular-matrix-bound LPL in an active form. Extracellular-matrix-bound LPL detached into medium containing triolein emulsion and/or serum, and was catalytically active after being released. Extracellular-matrix-bound LPL lost 30% of its activity following incubation at 37 degrees C for 4 h. in contrast to soluble LPL which lost 75% of its activity. It is plausible to conclude from these data that in vivo the subendothelial basement membrane, similarly to extracellular matrix, sequesters and stabilizers LPL secreted into the subendothelial space by non-endothelial cells, and thus may play an important role in determining the route of LPL from its site of synthesis to its site of action.
authors.affiliation: Lipid Research Laboratory, Department of Medicine B, Hadassah University Hospital, Jerusalem, Israel.
authors.forename: T
authors.initials: T
authors.lastname: Chajek-Shaul
authors.forename: G
authors.initials: G
authors.lastname: Friedman
authors.forename: G
authors.initials: G
authors.lastname: Bengtsson-Olivecrona
authors.forename: I
authors.initials: I
authors.lastname: Vlodavsky
authors.forename: R
authors.initials: R
authors.lastname: Bar-Shavit
mesh_terms: D000818:Animals; D002417:Cattle; D002478:Cells, Cultured; D003315:Cornea / Q000166:cytology / Q000378:metabolism; D004727:Endothelium / Q000166:cytology / Q000378:metabolism; D005109:Extracellular Matrix / Q000378:metabolism*; D006025:Glycosaminoglycans / Q000494:pharmacology; D006868:Hydrolysis; D008071:Lipoprotein Lipase / Q000378:metabolism*; D013917:Thrombin / Q000494:pharmacology; D014280:Triglycerides / Q000378:metabolism
chemical_list: D006025:Glycosaminoglycans; D014280:Triglycerides; D008071:Lipoprotein Lipase; D013917:Thrombin